The treatment of injuries or diseases affecting articular or elastic cartilage poses important unmet challenges to the medical community. The goal of this Phase I SBIR is to further the development of a method for the manufacture of a biologically-derived biomaterial that can maintain a pre-defined anatomical shape, while supporting the synthesis of a new tissue of a cartilage phenotype for the purpose of joint reconstruction or for reconstructive or plastic surgery. The novelty in this work entails the use o a biologically-derived material we have developed based solely on physical processing of allograft cartilage tissue. Our previous studies have demonstrated that this porous cartilage-derived matrix (CDM) can support, and even promote, chondrogenesis of both adipose-derived stem cells (ASCs) and mesenchymal stem cells (MSCs), resulting in a tissue that is biochemically and biomechanically similar to cartilage. To improve the shape retention properties of the biologically-derived biomaterial, CDM scaffolds will be cross-linked using chemical, UV light, or dehydrothermal processing. Following cross-linking of the matrix, the focus will be directed toward analysis of the proteome of the cross-linked CDM biomaterials to assess protein expression profiles between the different cross-linking treatments and also provide a baseline for future quality control during manufacture. The in vitro efficacy of the various cross-linked scaffolds seeded with ASCs or MSCs with respect to chondrogenic induction, shape maintenance, and mechanical integrity maintenance will also be evaluated. Measures of gene expression, tissue accumulation, and functional properties will be made using molecular, histologic, and biomechanical testing methods. Finally, a rabbit in vivo cartilage repair defect model will be used to assess the in vivo performance of the CDM by evaluating the regenerated tissue and overall joint tissue histologically and mechanically in comparison to the biochemical and biomechanical properties of native articular cartilage. Additionally, the overall toxicity and host immunological reaction to the CDM will be evaluated in this cartilage defect model. This scaffold technology will hopefully provide a novel means of developing tissue engineered constructs that are biomechanically functional at the time of creation and more easily integrated into host tissues in the body following surgical implantation. An improved level of biomechanical function will hopefully increase the level of success in the engineered repair of articular as well as elastic cartilage for applications in orthopaedic or plastic/reconstructive surgery. PUBLIC HEALTH RELEVANCE: The goal of this Phase I SBIR project is to develop a novel tissue engineered scaffold consisting of processed articular cartilage extracellular matrix that has been cross-linked so it can maintain a pre-defined anatomical shape. The ability of this porous, biologically active scaffold to support cartilaginous tissue formation will be tested using human adipose-derived or mesenchymal stem cells in long-term in vitro culture. Additionally, cartilaginous tissue growth and the host immunological reaction to the biomaterial will be monitored in an in vivo rabbit cartilage repair defect model. The ultimate goal of this project is o develop engineered tissues for treating cartilage defects in orthopaedic and/or reconstructive surgery applications.